摘要
将狂犬病病毒糖蛋白ｃＤＮＡＢｇｌⅡ（１．６７ｋｂ）片段分别正向插入ｐｍＭＴ－１ＢｇｌⅡ位点和ｐＭＴ０１０／Ａ＋ＢａｍＨＩ位点，构建重组质粒ｐｍＭＴ－Ｒｇｐ和ｐＭＴ０１０／Ａ＋－Ｒｇｐ，通过磷酸钙共沉淀法转染ＢＨＫ－２１细胞，经ＰＣＲ、打点杂交及ＥＬＩＳＡ检测，证明糖蛋白基因发生了整合并获得表达。以该表达载体给小鼠肌肉内注射２～３次，采其注射前、后双份血清，经ＥＬＩＳＡ检测证明，注射了表达载体的小鼠，血清中病毒特异性抗体明显升高，表明狂犬病病毒糖蛋白ｃＤＮＡ在小鼠体内表达后。
Expressing vectors, pmMT Rgp and pMT010/A + Rgp, were constructed by cloning rabies virus glycoprotein (Rgp) cDNA into downstream of the metallothionein promoter of pmMT 1 (murine) and pMT010/A + (ovine), respectively. Expression of the vectors in transfected BHK 21 cells was identified by dot hybridization of cellular RNAs with radio labelled probes of Rgp cDNA. The expressing vectors were then used to inoculate mice by injecting the vectors into quadricep muscles. Sera were obtained before the first injection and two weeks after the last injection. Specific antibodies against rabies virus were demonstrated to be present in the sera using Enzyme Linked Immune Sorbent Assay (ELISA) and Western blotting. It is clear that rabies virus glycoprotein cDNA were expressed and the mice were stimulated to develop humoral immune response.
出处
《中国兽医学报》
CAS
CSCD
1996年第4期321-326,共6页
Chinese Journal of Veterinary Science
基金
国家自然科学基金
